Morphine was nitrosated to 2-nitrosomorphine, and the morphine-2, 3-quinone-2-monopotassium oxime formed upon the addition of alakli to it was determined by polarograph under following conditions. By this means, it was found that the relationship id=CK is established within a range of 2×10-3 to 1×10-4 mole concentration. E1/2=1.13 volt vs (S.C.E.), kp=(I-1=C⋅kD-1⋅id-1)=1.113. The procedure follows: An aqueous solution of morphine was prepared in a range of 2×10-3 to 1×10-4 mole concentration. To 1cc. of this solution, 1.5cc. of N hydrochloric acid, and then 1.5cc. of 1 mole potassium nitrite were added, and allowed to stand at a room temperature for 1 minute to effect nitrosation. To this was added 1.5cc. of 20% potassium hydroxide solution, and its id, E1/2, and kp were determined by automatic recording between -0.4 to -1.3 V, at 25°±1°.
In order to establish quantitative determination of morphine preparations under the conditions investigated in the previous report, effect of the presence of impurities and coexisting substances were examined. As a result, a method for such determination has been proposed. This method was found to be effective when alkaloids such as various esters of morphine, dihydroxymorphine, atropine sulfate, and scopolamine hydrobromide, were present with morphine, but was not applicable with injections of morphine containing 0.3-0.5% phenol, and powders and tablets using sdsorption agents, such as aluminum silicate, as bulking agents. E1/2 of the quinone potassium oxime formed by the addition of alkali to the nitrosated phenol was found, to be -0.81 volts (S. C. E.).
A method for the determination of opium preparations was proposed as in the foregoing report. This followed in general the Stucki method modified by Ichikawa and Itoh. Morphine was extracted by a mixture of isopropyl alcohol and chloroform, and the extract was polarographically measured under conditions stipulated in the 1st Report. This method was found to be ineffective in determining marketed injections of opium preparations containing higher alcohols, such as glycerol or glycol.
Fungistatic and sporostatic effects of marketed antihistamines were tested against pathogenical fungi such as Trichophyton, Achorion, and Epidermophyton, by which Restamin and Anergen were found to be effective. With references to the works of others, it was seen that the antihistaminic compounds that are effective against fungi contain benzhydrol, phenothiazine, and benzylaniline group in their molecules. It was then found that these groups themselves exhibited far stronger fungistatic and spo-rostatic effects, sometimes surpassing 10 times the effect of the antihistamines. In other words, the efficacy of the antihistamines against fungi is not due to its antihist aminic activity but to the fungicidal action of their nuclear groups.
Fungistatic activities were tested with 24 kinds of bis- and triphenols against pathogenic fungi such as Trichophyton interdigitale and Achorion Schoenleini. In spite of their high antibacterial activities, only a few of the compounds were found to be effective in 1:8, 000 dilution, the others being ineffective at below 1:4, 000 dilution. The results show that there is no parallel relationship between bacteriostatic and fungistatic actions.
Using Lactobacillus fermenti, Lact. arabinosus and Streptococcus faecalis, tests were made of octopine, arcaine, and agmatine, the metabolic products of arginine, as to their arginine activity in arginine-free medium and inhibitory action in medium containing arginine. It was found that these three compounds do not act as arginine substitute or inhibitory in arginine determination. These results, together with the reports of Snell, seem to indicate that it would be possible to determine the content of arginine in the presence of ornithine, citrulline, agmatine, arcaine, and octopine, using Strept. faecalis.
Filter-type colorimeter, with a stable error of within 0.5%, was constructed using multiplier phototube MS-6S and RCA 931-A. The high voltage circuit is shown in Fig. 2, and their stability is shown in Figs. 4 and 5.
An automatic combustion apparatus for elemental analyses using a low frequency oscillation with Thyratron was used to test a few ideas on the speed of the movable furnace. From the results obtained, it was found that a constant speed of 1cm/min. for the driving area, and a few modifications in the fixed furnace and the combustion tube would result in a practical unit. These modifications would allow simplification of the driving unit and a shortening of the burning time.
Automatic temperature control method for heated mortar used for lead peroxide is described for laboratory use. The apparatus is an aluminum cylinder in whose wall several holes are punched and through which bimetal, combustion tube and nichrome wire are inserted. The movement of the contact point of bimetal is conducted by Thyratron, through which the main circuit is controlled. The aluminum cylinder is surface-treated with alumite so as to enable insertion of a live nichrome wire. The temperature variation inside the combustion tube, when this thermostatic furnace is used, was hardly detected by a thermometer.
The capacity of Anhydrone as a water absorbent in the analyses of carbon and hydrogen had been clearly demonstrated but it had the fault of being deliquescent and capable of contraction. In order to eliminate these shortcomings, the absorption tube was packed with Desichlora in the first half portion, and with Dehydrite in the latter portion, by which good results were obtained. This absorption tube, when the water absorptive power has reached a certain value, could be dehydrated at 140°, and repeatedly used.
By the elimination of lead peroxide and placing an absorbents of nitrogen oxides, there does not seem to occur any increase in the weight of water absorption tube due to the passage of nitrogen oxides. By the use of an absorbent tube filled with Desichlora and Dehydrite with glass-wool, no great obstruction was found in carrying out routine analyses. The use of cotton wool instead of glass-wool was found to give statistically a slight increase when there were a large formation of nitrogen oxides.
Formation of isonicotinic acid was examined by passing a mixed gas of 4-ethylpyridine and air, heated at 300-400°, over a catalyst, which included tin vanadate, vanadium pentoxide, and vanadium oxide containing a small amount of chromium oxide and tungsten oxide, with pumice stone as the carrier. The best result was obtained by the use of vanadium oxide containing a small amount of chromium oxide and tungsten oxide. The most suitable temperature was approximately 350° when the yield of isonicotinic acid was approximately 30% of the theoretical amount. No reaction products other than isonicotinic acid was detected.
By heating pyridine N-oxide and α-bromopyridine on a water bath, N-(α′-pyridyl)-α-pyridone was obtained. The same substance was also obtained by heating the sodium salt of α-pyridone and α-bromopyridine with copper dust at 200°. Similarly, N-(α′-pyridyl)-carbostyril was prepared from pyridine N-oxide and α-brolnoquinoline. This compound was also obtained by heating the sodium salt of carbostyril and α-bromopyridine with copper dust at 200°.
By the condensation of γ-aceto-γ-chloropropyl chloride (III), obtained by treating α-aceto-α-chlorobutyrolactone (II) with conc. hydrochloric acid in the presence of glacial acetic acid, with 2-methyl-4-amino-5-aminomethylpyrimidine (VI), ammonia, and carbon disulfide, 2′-methyl-2′-chlorotetrahydrofuryl-(3′) [2-methyl-4-aminopyrimidyl-(5)]-methyldithiocarbamate (VII) is obtained. Hydrogen chloride is easily liberated from (VII), and it transits easily to 2′-methyl-4′, 5′-dihydrof aryl-(3′) [2-methyl-4-aminopyrimidyl-(5)]-methyldithiocarbamate (VIII) by heating at 100°, or by treatment with alkali. (VIII) undergoes isomerization by treatment with diluted acids to 3-[2′-methyl-4′-aminopyrimidyl-(5′)]-methyl-4-methyl-5-β-hydroxyethylthiothiazolone-(2) (SB1) (I).
It was described in the previous paper that petroselidic acid was detected in the fruit oil of Anthriscus sylvestyis Hoffm., but that it was unknown whether the acid was contained, per se, in the fruit oil or whether it had been formed from petroselic acid during the extraction process. As a result of subsequent studies on the elaidination of petroselic acid, it was found that this acid underwent elaidination much more easily than oleic acid and, consequently, petroselic acid in the plant gradually underwent elaidination by the irradiation of ultraviolet light. However, elaidination was found not to occur by heating, saponification with alkalis, or by the action of diluted acids. Since the irradiation of ultraviolet light for an extended period failed to yield 100% petroselidic acid, it would be assumed that the fact that some crystals, m. p. 53°, of petroselidic acid had been obtained from the fruit oil of Anthriscus sylvestyis, shows that petroselidic acid is contained, per se, in the fruit oil, even in small quantities.
From 490g. of the fruit of Conioselinum univittatum Turcz., 55g. of crude fatty acids was obtained, and fractional distillation of their methyl ester (Cf. Table) gave four fractions. Fraction (A) and (B) yielded solid acids, that from the former being m. p. 61-62°, identical with palmitic acid, from and the latter, m. p. 55-58°, was identified by the paper chromatography of its hydroxamic acid as a mixture of palmitic acid and petroselic acid. Fraction (C) and (D) yielded solid acids of m. p. 31-32° and 32-33°, respectively, and the paper chromatography of their hydroxamic acids identified the former to be petroselic acid containing a minute amount of petroselidic acid, and the latter as petroselic acid. The liquid acids obtained from Fractions (C) and (D) were oxidized by the Hazula method and both yielded 6, 7-dihydroxystearic acid and 9, 10, 12, -13-tetrahydroxystearic acid. From these results, it was clarified that the fatty acids constituting the fruit oil was composed of palmitic, petroselic, petroselidic and linolic acids.
Studies were made on the inorganic constituent of ambergris obtained from the intestines of the sperm whale. The ether-insoluble matter (11.8%) of the ambergris is a dark brown powder, and contained some fragments of snout-like bone formations found in cuttle fish. Incineration of this powder yielded 16.3% of grayish white ash whose spectral analysis by the quartz spectrograph indicated the presence of the following. Group I - Ga, Fe*, K*, Mg, Na, (P), Si, Sr*, Zn; Group II - Al, Nb*, Cr*, Cu, La*, Mn, Ni*, Ti*, W* (where* denotes presence in minute amounts). The elements of the Group I were detected directly from the spectral lines, and those of the Group II were determined by comparing with spectrographs taken under the same conditions of the known compounds, such as aluminum oxide, columbite, chromium oxide, titanium oxide, and tungsten oxide, with that of ambergris ash. Phosphorus was confrmed as ammonium phosphomolybdate. It was assumed that a very minute amount of Ag, Co, Gd, and Hg were also present, although they were not determined. The quantitative analysis of the ash gave the following results. CaO 6.21%, MgO 9.88%, P2O5 4.65%, SiO2 6.02%.
2-Benzylaminopyrimidine was prepared by the application of propargyl aldehyde to benzylguanidine, and condensation with dimethylaminoethyl chloride gave N, N-dimethyl-N′-benzyl-N′-(2-pyrimidyl)-ethylenediamine. By a similar method, compounds containing methoxyl, bromine, chlorine, and fluorine in the para-position of the benzyl group were prepared. Application of cyanamide to N, N-dimethyl-N′-benzylethylenediamine gave 1-dimethylaminoethyl-1-benzylguanidine, reaction of propargyl aldehyde to which yielded N, N-dimethyl-N′-benzyl-N′-(2-pyrimidyl)-ethylenediamine. Pharmacological tests revealed that the compound possessing fluorine in the para-position of the benzyl group had the most strongest antihistamine acitivity.
By the condensation of 2-benzylthiazoles (III), obtained by the application of dichloroether to phenylthioacetamides (II), and dimethyl aminoethyl chloride or N-piperidylethyl chloride, thiazolylpropylamine derivatives (IV and V) were prepared. These were also obtained by the application of dichloroether to thiobutyroamide derivatives (VIII and IX), obtained by the reaction of hydrogen sulfide to α-phenyl-γ-dimethylaminobutyronitrile (VI) or α-phenyl-γ-(N-piperidyl)-butyronitrile (VII). Of these compounds, N, N-dimethyl-3-phenyl-3-(2-thiazolyl)-propylamine derivatives possessing a halogen atom in the para-position in the phenyl ring have excellent antihistamine properties. It was also found that the efficacy increased in the order of fluorine, chlorine, and bromine.
Attempts to obtain the lactone of α-(2-hydroxy-3-ketocyclohexyl)-propionic acid (II) by the hydration of the double bond of diethyl (3-ketocyclohexen-(1)-yl)-methylmalonate (I) were unsuccessful and, therefore, the double bond in (I) was hydroxylated in aqueous solution of potassium permanganate, saponif ied, and heated in acetic anhydride, by which the following three unsaturated lactones, closely analogous to (II), were obtained: Lactones of α-(2-hydroxy-3-acetoxycyclohexen-(2)-ylidene)-propionic acid (XII), of α-(2-hydroxy-3-acetoxycyclohexadien-(1, 3)-yl)-propionic acid (XIII), and of α-(2-hydroxycyclohexen-(1)-yl)-propionic acid (XIV). Other allied compounds are also described.
As a reagent for testing the properties of oxine derivatives in inducing diabetes, oxine homologs and their azo derivatives were synthesized, and the following facts were found: 1) Compounds which have better inducing properties than oxine: 8-Hydroxyquinaldine, 2, 4-dimethyl-8-hydroxyquinoline, and 5-(phenyl)-azo, 5-(p-tolyl)-azo, and 5-(p-hydroxyphenyl)-azo derivatives of oxine and 8-hydroxyquinaldine. 2) Compounds which do not induce diabetes: 5-(p-Sulfophenyl)- and 5-(m-carboxy-phenyl)-azo-8-hydroxyquinoline, 2, 4-dihydroxyquinaldine, xanthurenic acid, 8-methoxy-quinoline, and 8-methoxyquinaldine. 3) Compounds which induce diabetes and which selectively dyes the Langerhans' island cells: 5-(p-Hydroxyphenyl)-azo-8-hydroxyquinoline and 5-(p-hydroxypherlyl)-azo-8-hydroxyquinaldine. The syntheses of xanthurenic acid and 2, 4-dihydroxyquinaldine gave slightly different melting points of the intermediate of the former and that of the latter from those described by Mebane and Wiederkehr.
Equine parotic gland is fairly different from bovine parotic gland, both in outward appearance and in properties. The pH 5.4-precipitate obtained from equine gland is larger in yield than that from bovine gland, but its effect is far weaker. The 12% (NH4)2SO4-fraction, obtained by the purification of the equine pH 5.4-precipitate by fractional precipitation with ammonium sulfate, could not be induced to crystallize, remaining granular. The effect of this fraction is much weaker than the 12.1% (NH4)2SO4-fraction from the bovine gland, and was found to be still rather impure from the results of electrophoretic analysis. The mobility of the active fraction obtained from the equine parotic gland is much larger than that of the active fraction from the bovine gland. The presence of a substance in the equine parotic gland which lowers the calcium level of rabbit serum is similar to the case in bovine gland, but its properties other than such physiological action, are slightly different from those of the bovine gland.
Parotin not only lowers the calcium level of rabbit serum but it also remarkably increases the phosphorus level, especially that of inorganic phosphorus. The fractions corresponding to parotin, obtained from bovine thymus glands, spleen, and heart, and the bovine serum albumin and globulin, also increase the serum phosphorus level but the rate of increase is extremely small. Their action on phosphorus, as is the case on calcium, is of different type from that of parotin.
A fraction (precipitating at the isoelectric point), corresponding to crude parotin, obtained from the aqueous extracts of bovine liver, thymus, and spleen, and the albumin and globulin from bovine serum, also showed activity of decreasing the calcium level of rabbit serum, but its mode of action differs from that of parotin (Ogata, Ito, Nozaki, and Okabe: This Journal, 64, 114 (1944): Ogata, Ito: This Journal.. 64, 332 (1944)). By a similar procedure, pH 5.7-precipitate and pH 5.4-precipitate were obtained from the aqueous extract of bovine heart muscles. The pH 5.7-precipitate also possesses an action like parotin of lowering serum calcium level, but in higher dosage (30mg./2kg. wt.), it acts reversely to increase the calcium level. Solubilities of these various principles in water of various pH, and the manner of denaturation when boiled with water of various pH values, were examined. Quantitative determination of total nitrogen, amino nitrogen, phosphorus, and sulfur contents of these products showed that their properties differed from those of parotin except the pH 5.7-precipitate from bovine heart which was very similar in its properties to parotin.
β-Dimethylaminoethyl thiobenzhydryl ether hydrochloride has been found to be effective as an antihistamine from clinical tests. Following compounds, belonging to the same series of compounds, were prepared, of which, the first one had been synthesized by Rieveshl: β-Dimethylaminoethyl p-methylthiobenzhydryl, p-chlorothiobenzhydryl, and p, p′-dichlorothiobenzhydryl ether hydrochloride.
In order to examine the effect of chemical structure on antibacterial activity, some pyridine carboxylic acid derivatives shown in the Table were prepared. The results of in vitro tests against the so-called “Mycobacterium tuberculosis A. T. C. C. No. 607” are also given in the Table. Of the three isomers of acid hydrazide, the one of γ-substituent, i.e. isonicotinic acid hydrazide (INAH), alone showed excellent antibacterial action, which decreased when the compound was derived to its N-oxide. During the preparation of above samples, ethyl picolinate was found to undergo saponification by treatment with hydrogen peroxide and glacial acetic acid, forming picolinic acid N-oxide. With the others, esters were derived to their N-oxides by treatment with hydrogen peroxide and glacial acetic acid, then led to acid amides and acid hydrazides with aqueous ammonia and hydrazine hydrate, respectively.
It has been maintained that the essential oil (Chenopodium Oil) obtained from Chenopodium ambrosioides L. var. anthelminticum A. Gray cultivated in the cooler districts of Japan is not quite as good a quality as that from the plants cultivated in the warmer climates, but no experimental evidence for it has ever been offered. The plant was experimentally cultivated in the highlands of the Nagano Prefecture, the Tohoku Districts, and in the Hokkaido, which are in a cooler region, and distillation tests and determination of ascaridol were carried out on the crop. The results obtained were as follows: 1) Table I shows the growth, yields of crop and fruits, content of the essential oil, and the content of ascaridol. Tables II and III respectively show the atmospheric temperature and the rainfall in these districts. 2) The growth, especially in the first stage, is retarded in cooler regions, but the yields of crop and of fruits were not necessarily less than those from the warm region. 3) The yield of the oil and the content of ascaridol were also not particularly less in the crop from the cooler region compared to those from the warm region.
Particulars obtained for 3-oximino-4-oxohomocamphor, m. p. 112°, described in the previous report, are as follows: 1) Besides forming colored complex salts with potassium and ferrous ions, it also forms colored salts with other metallic ions. 2) It gives camphoric acid and camphoric anhydride in a good yield with reagents that induces the Beckmann rearrangement. It was also confirmed that, in the case of conc. sulfuric acid, carbon monoxide is generated at the same time, so that its reaction mechanism must belong to the second type Beckmann rearrangement, and the formation of camphoric acid monocyanide (VIII) as an unstable intermediate, was assumed. 3) Of the two possible structures of (Va) and (Vb), (Va) was assumed to be more appropriate for 3-oximino-4-oxohomocamphor.
Nitration of benzaldazine with sulfuric and nitric acids gave o, o′-dinitrobenzaldazine and p, p′-dinitrobenzaldazine, but their yields were very poor, the majority undergoing decomposition. However, this has definitely shown that nitro substitutions occurred in both the ortho and the para positions. Nitration of benzaldazine with acetic anhydride and nitric acid yielded benzaldehyde and 4-benzalamino-3, 5-diphenyhl-1, 2, 4-triazole, m. p. 205.5-206.5° (picrate, 169-170°), but no nitro derivatives. Benzaldazine refused to be nitrated with nitric acid in glacial acetic acid, the starting material being recovered. It was assumed that the activation of ortho and para positions during the nitration of benzaldazine with sulfuric and nitric acids was due probably to the electron displacement by the liberation of a proton, which took up an unstable coordination against a weak base by the effect of sulfuric acid.
Oxidation of benzaldazine in ether solution with perphthalic acid gives monobenzyl-phthalate, while oxidation with hydrogen peroxide in glacial acetic acid gives benzyl-acetate, Benzaldazine hardly undergoes oxidation in neutral medium The mechanism of the formation of benzylesters by the oxidation of benzaldazine was assumed to be due to the formation of phenyl diazomethane as an intermediate during the reaction.
In the preparation of 2-thiocyano-3-nitropyridine by the condensation of 2-chloro-3-nitropyridine and potassium thiocyanate, the addition of activated copper dust results in an increased yield. Respective application of benzoyl chloride and p-nitrobenzoyl chloride to 2-mercapto-3-amino-6-chloropyridine gave pyrido-2, 3:5′, 4′-thiazole derivatives. Application of p-nitrosodimethylaniline and p-nitrosodiethylaniline to 2′-methyl-6-chloropyrido-2, 3:5′, 4′-thiazole gave the corresponding azomethine compounds.
2-Dimethylamino-6-methoxybenzothiazole was changed to its 6-hydroxy derivative by boiling with hydriodic acid (d=1.7), and the respective condensation of isopropyl iodide, butyl bromide, and isoamyl bromide to the hydroxy derivative gave the corresponding 6-alcoxy derivatives. By the condensation of aromatic aldehydes to 2-allyl-mercapto (chloro)-6-aminobenzothiazole, 16 kinds of Schiff's bases were prepared. Some of these bases showed strong tuberculostatic activity, in vitro.
1) Reaction between 1-phenylthiosemicarbazide and various metallic ions was examined and it was found that the compound gave sharp coloration with copper, silver, gold, zinc, cadmium, mercury, bismuth, iron, cobalt, nickel, and platinum ions. 2) It was found that the color reaction with cupric ion was especially sharp and that the isoamyl alcohol solution of this reagent was able to detect 2×10-8g./cc. of cupric ion. 3) Reagent test papers were prepared and the limit of the detection of copper ion and the limit ratio, in the presence of other metallic ions, were examined with these papers. 4) The use of this reagent as the color developer in the paper chromatography enabled the concurrent separatory detection of various metallic ions.
Physical and chemical constants of parotin (Fig. 4), which is eletrophoretically almost homogeneous, were determined (Tables I and II). The stability of the aqueous solution of parotin was also examined (Table II).
Various chemical reagents were applied on parotin in order to examine the essential groups responsible for the appearance of its effect as the lowering of serum calcium level. This effect was found to either decrease or disappear when the free amino radical in parotin was fixed or shielded by the ketene, nitrous acid, or formaldehyde, or when the thyrosine-phenyl ring was substituted by iodine. Treatment of parotin with 40 volumes of cysteine hydrochloride, in order to reduce the -S-S- bond in its molecule, gave no effect on the calcium lowering activity, but the treatment with 20 volumes of thioglycollic acid was found to decrease the effect (at level of significance 7.3%). From these experiments, it is assumed that the free amino radical, thyrosinephenyl ring, and the -S-S- bond are responsible for the appearance of the specific effect in parotin.
It was found that, taking the parotin test of the lowering of calcium level in rabbit serum as the criterion, treatment of parotin with 3.0 and 6.66 moles urea solution (pH 7.0) resulted in reversible denaturation (Table I), while the treatment with 6.0 moles guanidine hydrochloride solution (pH 7.0) resulted in irreversible denaturation (Table II). The mean lowering rate of serum calcium level, when 4cc. per 2kg. wt. of physiological saline solution is intravenously injected, into rabbits (5 rabbits were used), is 5.61%, while it is 2.13% when 1cc. per 2kg. wt. is injected (Table IV). It was assumed that if the amount of sample intravenously injected into rabbits is around 1cc. per 2kg. of weight, such injection has practically no effect on the lowering rate of serum calcium level.
The parotin preparation of 93% purity (electrophoretically analyzed. Fig. 1) gives negative pentose reaction (by phloroglucinol-hydrochloric acid and orcinol-hydrochloric acid), but gives positive carbazole-sulfuric acid reaction. The absorption curve of the parotin preparation by the latter reaction greatly differs from those of mannose, galactose and glucose (Fig. 2), but the curve for parotin hydrolyzate is very similar to those of the sugars (Fig. 3). In the absorption curves shown in Fig. 3, the E520/E420 for parotin hydrolyzate is 0.73, while those of galactose is 0.99 and mannose, 0.76 (Gurin: galactose 1.10, mannose 0.68), showing its value approximating that of mannose. However, from the color tone, absorption curve, and the E520/E420 value, parotin hydrolyzate does not seem to contain glucose.
Based on the assumption that the sugar in parotin is mannose, the amount of sugar contained in parotin of approximately 93% purity was 0.73%. Less pure parotin, recovered from the precipitation mother liquor, obtained during the purification of parotin, contained 1.78% of sugar. From these results, it is assumed that the sugar content of parotin itself is in a range of 0.79 to 0.48%. It is also assumed that hexos amine is either nonexistent or is contained in a minute amount, and that a very minute amount of pyrrole dye is contained in parotin preparation.
Bovine submaxillary gland contains a substance which remarkably lowers the serum phosphorus level. This substance is present in a water-soluble fraction, and precipitates at pH 4.5. It is unstable to heat, and seems to be affected by alcohol. Its solubility to water of various pH and coagulation reactions are similar to those of parotin, a substanc eobtained from the bovine parotic gland and possessing the action of decreasing serum calcium level.
Some isomers of di- and tri-hydroxybenzoic acids were prepared to test their activities on tubercle bacilli. Of the compounds prepared, 2, 3, 6-trihydroxybenzoic acid is unknown in literature, whose preparation from 2, 6-dihydroxybenzoic acid by means of alkaline persulfate oxidation was found to yield no crystalline product. The methylation of the crude product with dimethyl sulfate finally yielded 2, 3, 6-trimethoxybenzoic acid.
An acidic saponin was isolated in a pure crystalline state from the seed of Thea sinensis L., cultivated in Shizuoka Prefecture, and was designated thea-saponin. It melted at 224-225° (decomp.) (corr.), [α]D19=+37.6° (dil. EtOH); hemolytic index, ca. 100, 000 (with rabbit blood). From the results of analytical data and the sugar components, the formula of C57H90O26 was given for this triterpenoid saponin. Alkaline hydrolysis of thea-saponin converted it into thea-prosapogenol A, C52H84O25, m. p. 227°, [α]D19=+43.6° (dil. EtOH)., with liberation of one mole of angelic acid.
Thea-saponin was finally hydrolyzed to a monoacidic sapogenin, thea-sapogenol, m.p. 301-302.5°, [α]D19=+77.3° (EtOH), four sugar components, and one mole of angelic acid. The four sugars were identified, both by paper chromatography and as osazone derivatives. These results can be summarized as follows: Thea-saponin C57H90O26+5H2O=Thea-sapogenol C30H50O6+glucuronic C6H10O7 acid+galactose C6H12O6+arabinose C5H10O5+xylose C5H10O5+angelic C5H8O2 acid